Skip to main content
SpringerLink
Log in

Neuroprotection of Interleukin-6 Against NMDA-induced Neurotoxicity is Mediated by JAK/STAT3, MAPK/ERK, and PI3K/AKT Signaling Pathways

  • Original Research
  • Published:
Cellular and Molecular Neurobiology Aims and scope Submit manuscript

Abstract

We have previously shown that interleukin-6 (IL-6) has neuroprotective effect against N-methyl-d-aspartate (NMDA)-induced excitotoxicity. The current study aimed to reveal signal transduction pathways involved in the IL-6 neuroprotection. Cerebellar granule neurons (CGNs) from postnatal 8-day infant rats were exposed to IL-6 (120 ng/ml) for 8 days and stimulated with NMDA (100 μM) for 15 or 30 min. Dynamic intracellular Ca2+ fluorescence intensity, cytosolic Ca2+-dependent phospholipase A2 (cPLA2) expression, and apoptosis and necrosis in cultured CGNs were measured by laser scanning confocal microscope, real-time PCR and Western blot, and annexin V-FITC/propidium iodide staining, respectively. NMDA stimulation of neurons evoked an intracellular Ca2+ overload, an upregulated expression of cPLA2, and an increase in cell death. Chronic IL-6 exposure prevented the NMDA-evoked neuronal Ca2+ overload, cPLA2 expression upregulation, and apoptosis and necrosis. Anti-gp130 monoclonal antibody (mAb), a blocker of gp130 that is a 130-kDa signal-transducing β-subunit of IL-6 receptor complex, blocked these effects of IL-6 preventing NMDA neurotoxicity. AG490, PD98059, or LY294002, inhibitors specific for the intracellular signals, JAK, MAPK, and PI3K, respectively, partially blocked these IL-6 neuroprotective effects. Phosphorylation levels of STAT3, ERK1/2, and AKT, the downstream proteins for these enzymes of JAK, MAPK, and PI3K, respectively, were elevated by IL-6 pretreatment. The enhanced activation of STAT3, ERK1/2, and AKT by IL-6 was abolished by AG490, PD98059, and LY294002, respectively. Anti-gp130 mAb attenuated the activation of all the three detected signaling molecules. The present findings suggest that IL-6 neuroprotection is jointly mediated by the cellular signal transduction pathways, gp130-JAK-STAT3, gp130-MAPK-ERK, and gp130-PI3K-AKT.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

References

  • Ali C, Nicole O, Docagne F, Lesne S, MacKenzie ET, Nouvelot A, Buisson A, Vivien D (2000) Ischemia-induced interleukin-6 as a potential endogenous neuroprotective cytokine against NMDA receptor-mediated excitotoxicity in the brain. J Cereb Blood Flow Metab 20:956–966

    Article  PubMed  CAS  Google Scholar 

  • Arundine M, Tymianski M (2003) Molecular mechanisms of calcium-dependent neurodegeneration in excitotoxicity. Cell Calcium 34:325–337

    Article  PubMed  CAS  Google Scholar 

  • Baker DG, Ekhator NN, Kasckow JW, Hill KK, Zoumakis E, Dashevsky BA, Chrousos GP, Geracioti TD Jr (2001) Plasma and cerebrospinal fluid interleukin-6 concentrations in posttraumatic stress disorder. NeuroImmunoModulation 9:209–217

    Article  PubMed  CAS  Google Scholar 

  • Baranzini SE, Elfstrom C, Chang SY, Butunoi C, Murray R, Higuchi R, Oksenberg JR (2000) Transcriptional analysis of multiple sclerosis brain lesions reveals a complex pattern of cytokine expression. J Immunol 165:6576–6582

    PubMed  CAS  Google Scholar 

  • Bauer J, Strauss S, Volk B, Berger M (1991) IL-6-mediated events in Alzheimer’s disease pathology. Immunol Today 12:422

    Article  PubMed  CAS  Google Scholar 

  • Bonventre JV (1996) Roles of phospholipases A2 in brain cell and tissue injury associated with ischemia and excitotoxicity. J Lipid Mediat Cell Signall 14:15–23

    Article  CAS  Google Scholar 

  • Burke JE, Dennis EA (2009) Phospholipase A2 structure/function, mechanism, and signaling. J Lipid Res 50:S237–S242

    Article  PubMed  Google Scholar 

  • Campbell IL, Abraham CR, Masliah E, Kemper P, Inglis JD, Oldstone MB, Mucke L (1993) Neurologic disease induced in transgenic mice by cerebral overexpression of interleukin 6. Proc Natl Acad Sci USA 90:10061–10065

    Article  PubMed  CAS  Google Scholar 

  • Chang JY, Tsai PF (2009) IL-6 release from mouse glia caused by MeHg requires cytosolic phospholipase A2 activation. Neurosci Lett 461:85–89

    Article  PubMed  CAS  Google Scholar 

  • Cucchiaroni ML, Viscomi MT, Bernardi G, Molinari M, Guatteo E, Mercuri NB (2010) Metabotropic glutamate receptor 1 mediates the electrophysiological and toxic actions of the cycad derivative beta-N-methylamino-l-alanine on substantia nigra pars compacta DAergic neurons. J Neurosci 30:5176–5188

    Article  PubMed  CAS  Google Scholar 

  • D’ Arcangelo G, Tancredi V, Onofri F, D’ Antuono M, Giovedi S, Benfenati F (2000) Interleukin-6 inhibits neurotransmitter release and the spread of excitation in the rat cerebral cortex. Eur J Neurosci 12:1241–1252

    Article  Google Scholar 

  • Farooqui AQA, Ong WY, Horrocks LA (2006) Inhibitors of brain phospholipase A2 activity: their neuropharmacological effects and therapeutic importance for the treatment of neurologic disorders. Pharmacol Rev 58:591–620

    Article  PubMed  CAS  Google Scholar 

  • Fernández-Sánchez MT, Novelli A (1993) Basic fibroblast growth factor protects cerebellar neurons in primary culture from NMDA and non-NMDA receptor mediated neurotoxicity. FEBS Lett 235:124–131

    Article  Google Scholar 

  • Frei K, Fredrikson S, Fontana A, Link H (1991) Interleukin-6 is elevated in plasma in multiple sclerosis. J Neuroimmunol 31:147–153

    Article  PubMed  CAS  Google Scholar 

  • Fukada T, Hibi M, Yamanaka Y, Takahashi-Tezuka M, Fujitani Y, Yamaguchi T, Nakajima K, Hirano T (1996) Two signals are necessary for cell proliferation induced by a cytokine receptor gp130: involvement of STAT3 in anti-apoptosis. Immunity 5:449–460

    Article  PubMed  CAS  Google Scholar 

  • Gadient RA, Otten U (1993) Differential expression of interleukin-6 (IL-6) and interleukin-6 receptor (IL-6R) mRNAs in rat hypothalamus. Neurosci Lett 153:13–16

    Article  PubMed  CAS  Google Scholar 

  • Gadient RA, Otten U (1994) Identification of interleukin-6 (IL-6)-expressing neurons in the cerebellum and hippocampus of normal adult rats. Neurosci Lett 182:243–246

    Article  PubMed  CAS  Google Scholar 

  • Gentile MT, Reccia MG, Sorrentino PP, Vitale E, Sorrentino G, Puca AA, Colucci-D’Amato L (2012) Role of cytosolic calcium-dependent phospholipase A2 in Alzheimer’s disease pathogenesis. Mol Neurobiol 45:596–604

    Article  PubMed  CAS  Google Scholar 

  • Gruol DL, Nelson TE (2005) Purkinje neuron physiology is altered by the inflammatory factor interleukin-6. Cerebellum 4:198–205

    Article  PubMed  CAS  Google Scholar 

  • Ha BK, King JS (2000) Localization of gp130 in the developing and adult mouse cerebellum. J Chem Neuroanat 19:129–141

    Article  PubMed  CAS  Google Scholar 

  • Hibi M, Hirano T (2000) Gab-family adapter molecules in signal transduction of cytokine and growth factor receptors, and T and B cell antigen receptors. Leuk Lymphoma 37:299–307

    PubMed  CAS  Google Scholar 

  • Inomata Y, Hirata A, Yonemura N, Koga T, Kido N, Tanihara H (2003) Neuroprotective effects of interleukin-6 on NMDA-induced rat retinal damage. Biochem Biophys Res Commun 302:226–232

    Article  PubMed  CAS  Google Scholar 

  • Kamimura D, Ishihara K, Hirano T (2003) IL-6 signal transduction and its physiological roles: the signal orchestration model. Rev Physiol Biochem Pharmacol 149:1–38

    Article  PubMed  CAS  Google Scholar 

  • Kriem B, Sponne I, Fifre A, Malaplate-Armand C, Lozac’h-Pillot K, Koziel V, Yen-Potin FT, Bihain B, Oster T, Jean-Luc Olivier, Pillot T (2005) Cytosolic phospholipase A2 mediates neuronal apoptosis induced by soluble oligomers of the amyloid-β peptide. FASEB J 19:85–87

    PubMed  CAS  Google Scholar 

  • Kushima Y, Hama T, Hatanaka H (1992) Interleukin-6 as a neurotrophic factor for promoting the survival of cultured catecholaminergic neurons in a chemically defined medium from fetal and postnatal rat midbrains. Neurosci Res 13:267–280

    Article  PubMed  CAS  Google Scholar 

  • Liu Z, Qiu YH, Li B, Ma SH, Peng YP (2011) Neuroprotection of interleukin-6 against NMDA-induced apoptosis and its signal-transduction mechanisms. Neurotox Res 19:484–495

    Article  PubMed  CAS  Google Scholar 

  • Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using Real-Time quantitative PCR and the 2–44Ct method. Methods 25:402–408

    Article  PubMed  CAS  Google Scholar 

  • Merrill JE, Chen IS (1991) HIV-1, macrophages, glial ells, and cytokines in AIDS nervous system disease. FASEB J 5:2391–2397

    PubMed  CAS  Google Scholar 

  • Nelson TE, Olde Engberink A, Hernandez R, Puro A, Huitron-Resendiz S, Hao C, De Graan PN, Gruol DL (2012) Altered synaptic transmission in the hippocampus of transgenic mice with enhanced central nervous systems expression of interleukin-6. Brain Behav Immun 26:959–971

    Article  PubMed  CAS  Google Scholar 

  • Peng YP, Qiu YH, Lu JH, Wang JJ (2005) Interleukin-6 protects cultured cerebellar granule neurons against glutamate-induced neurotoxicity. Neurosci Lett 374:192–196

    Article  PubMed  CAS  Google Scholar 

  • Pizzi M, Sarnico I, Boroni F, Benarese M, Dreano M, Garotta G, Valerio A, Spano P (2004) Prevention of neuron and oligodendrocyte degeneration by interleukin-6 (IL-6) and IL-6 receptor/IL-6 fusion protein in organotypic hippocampal slices. Mol Cell Neurosci 25:301–311

    Article  PubMed  CAS  Google Scholar 

  • Qiu Z, Gruol DL (2003) Interleukin-6, beta-amyloid peptide and NMDA interactions in rat cortical neurons. J Neuroimmunol 139:51–57

    Article  PubMed  CAS  Google Scholar 

  • Sharp JD, White DL (1993) Cytosolic PLA2: mRNA levels and potential for transcriptional regulation. J Lipid Mediat 8:183–189

    PubMed  CAS  Google Scholar 

  • Shelat PB, Chalimoniuk M, Wang JH, Strosznajder JB, Lee JC, Sun AY, Simonyi A, Sun GY (2008) Amyloid beta peptide and NMDA induce ROS from NADPH oxidase and AA release from cytosolic phospholipase A2 in cortical neurons. J Neurochem 106:45–55

    Article  PubMed  CAS  Google Scholar 

  • Spooren A, Kolmus K, Laureys G, Clinckers R, De Keyser J, Haegeman G, Gerlo S (2011) Interleukin-6, a mental cytokine. Brain Res Rev 67:157–183

    Article  PubMed  CAS  Google Scholar 

  • Stahl N, Farruggella TJ, Boulton TG, Zhong Z, Darnell JE Jr, Yancopoulos GD (1995) Choice of STATs and other substrates specified by modular tyrosine-based motifs in cytokine receptors. Science 267:1349–1353

    Article  PubMed  CAS  Google Scholar 

  • Sun XM, Lu JH, Qiu YH, Liu Z, Wang XQ, Peng YP (2011) Interleukin-6 reduces NMDA-induced Ca2+ overload via prevention of Ca2+ release from intracellular store. Int J Neurosci 121:423–429

    Article  PubMed  CAS  Google Scholar 

  • Tran HY, Shin EJ, Saito K, Nguyen XK, Chung YH, Jeong JH, Bach JH, Park DH, Yamada K, Nabeshima T, Yoneda Y, Kim HC (2012) Protective potential of IL-6 against trimethyltin-induced neurotoxicity in vivo. Free Radic Biol Med 52:1159–1174

    Article  PubMed  CAS  Google Scholar 

  • Van Wagoner NJ, Benveniste EN (1999) Interleukin-6 expression and regulation in astrocytes. J Neuroimmunol 100:124–139

    Article  PubMed  Google Scholar 

  • Wang XQ, Peng YP, Lu JH, Cao BB, Qiu YH (2009) Neuroprotection of interleukin-6 against NMDA attack and its signal transduction by JAK and MAPK. Neurosci Lett 450:122–126

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgments

We wish to thank Dr. M. Kerry O’Banion, University of Rochester, for editing this manuscript. This work was supported by Grants BK2010278 and BK2011386 from the Natural Science Foundation of Jiangsu Province of China; K2010047, BK2012014, BK2012015, and CP12012003 from the Nantong Applied Research Program of China; and a Project funded by the Priority Academic Program Development of Jiangsu Higher Education Institutions.

Conflict of interest

The authors declare that they have no conflict of interest.

Author information

Authors and Affiliations

  1. Department of Physiology, School of Medicine, Nantong University, 19 Qixiu Road, Nantong, 226001, China

    Xiao-Xia Fang, Xiao-Lin Jiang, Xin-Hua Han, Yu-Ping Peng & Yi-Hua Qiu

Authors
  1. Xiao-Xia Fang
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Xiao-Lin Jiang
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Xin-Hua Han
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Yu-Ping Peng
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Yi-Hua Qiu
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding authors

Correspondence to Yu-Ping Peng or Yi-Hua Qiu.

Additional information

Xiao-Xia Fang and Xiao-Lin Jiang contributed equally to this study.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Fang, XX., Jiang, XL., Han, XH. et al. Neuroprotection of Interleukin-6 Against NMDA-induced Neurotoxicity is Mediated by JAK/STAT3, MAPK/ERK, and PI3K/AKT Signaling Pathways. Cell Mol Neurobiol 33, 241–251 (2013). https://doi.org/10.1007/s10571-012-9891-6

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10571-012-9891-6

Keywords

Search

Navigation